Structural GenomiX, Inc., Albert Einstein College of Medicine, the Department of Biology at Brookhaven National Laboratory, Columbia University, Sloan Kettering Institute, and the University of California at San Francisco comprise the New York Structural Genomics Research Consortium (NYSGXRC). This U54 application proposes to establish and operate a Large Scale Center for Phase 2 of the Protein Structure Initiative (PSI-2), as specified in RFA-GM-05-001. Together, members of the NYSGXRC will establish and operate a fully-integrated, high-throughput center for the following activities: (i) protein family classification and target selection, (ii) protein expression and solubility testing, (iii) E. coli and baculovirus fermentation, (iv) purification and biophysical characterization, (v) crystallization, (vi) synchrotron X-ray diffraction data collection, (vii) X-ray phase determination via molecular replacement, multi- and single-wavelength anomalous dispersion measurements, and isomorphous replacement, (viii) model building and refinement, (ix) PDB deposition of atomic coordinates and experimental structure factors, (x) comparative protein structure modeling, and (xi) functional annotation and dissemination of results. Targets for this effort will be derive from three sources, including the PSI-2 Centralized Target Selection Committee (ca. 70% of capacity), an NYSGXRC biomedical research program focused on the phosphatase superfamily (ca. 15% of capacity), and targets solicited from the larger scientific community (ca. 15% of capacity). A program of technology development will also be undertaken to both increase efficiency/throughput and accelerate structure determination for challenging, multi-domain eukaryotic proteins. At peak production, the NYSGXRC will be capable of determining 200 or more structures annually at an average cost approaching $50,000-60,000/structrure for proteins that can be expressed in E. coli. The long-term goal of the NYSGRC is to contribute substantially to the 5,000-10,000 structure objective of PSI-2.